QUAPO : Quantitative Analysis of Pooling in High-Throughput Drug - - PowerPoint PPT Presentation

QUAPO : Quantitative Analysis of Pooling in High-Throughput Drug Screening

Raghu Kainkaryam

Systems Biology Group University of Michigan (joint work with Anna Gilbert, Paul Shearer and Peter Woolf) March 27, 2009 DIMACS/DyDAn Workshop on Streaming, Coding & Compressive Sensing

Motivation Pooling in HTS QUAPO Challenges Summary

Talk Outline

1 Motivation Drug Discovery HTS 2 Pooling in HTS Group Testing 3 QUAPO Compressive Sensing Results 4 Challenges Practical Challenges 5 Summary Take Home Points

Motivation Pooling in HTS QUAPO Challenges Summary

Drug Discovery Funnel

Motivation Pooling in HTS QUAPO Challenges Summary

Drug Discovery Cost

• Approx. Cost ∼ $800 million to bring a new drug to market1

New drug = New Chemical Entity Each year, worldwide, only about 26 such drugs enter the market Millions of chemical compounds are tested to find them

1includes the cost of all drug development which did not result in a new drug

Motivation Pooling in HTS QUAPO Challenges Summary

High-Throughput Screening (HTS) First step in drug discovery is High-Throughput Screening (HTS).

Motivation Pooling in HTS QUAPO Challenges Summary

ABC of HTS Automation & high-throughput achieved through robotic liquid handling Biological Assay – Typically a biochemical binding event detected by an optical signal Chemical Library – thousands to millions of chemical compounds, available in pre-configured plates. Hit Rate – number of active compounds found in a screen (0.01 – 10%)

Motivation Pooling in HTS QUAPO Challenges Summary

Pooling in HTS Comparison of one compound, one well and pooled HTS.

Motivation Pooling in HTS QUAPO Challenges Summary

Multiple Items & Noisy Tests Unique boolean tagging does not work when multiple active compounds

• r testing errors occur.

Motivation Pooling in HTS QUAPO Challenges Summary

Group Testing Problem : Create pooling strategy that reduces tests, guarantees identification and corrects errors in testing.

Solution : Group Testing 2

For n compound library With at most k active With at most E testing errors Design pooling strategy to guarantee the identification of k actives Design a decoding algorithm which works in the presence of E errors

2which means Compressive Sensing is around the corner

Motivation Pooling in HTS QUAPO Challenges Summary

Pooling Design Example: Shifted Transversal Design (STD) of N.Thierry Mieg 3 for n = 25, k = 2, E = 1.

3shown to be equivalent to R. DeVore’s Deterministic Construction (2007)

Motivation Pooling in HTS QUAPO Challenges Summary

Decoding Algorithm

Choose a cut-off to reduce measurements to binary (hit or miss).4

4figures from K. & Woolf, Curr. Op. in Drug Disc. & Dev, in press 2009

Motivation Pooling in HTS QUAPO Challenges Summary

Quantitative Analysis of Pooling Quantitative information is present in measurements. Binary binning of data introduces false positive and false negative testing errors. Hard to choose cut-off for pooled measurements.

Motivation Pooling in HTS QUAPO Challenges Summary

Compressive Sensing in HTS Quantitative Analysis of Pooling is possible via Compressive Sensing. It is sparse but is it linear?

Motivation Pooling in HTS QUAPO Challenges Summary

Biochemical Model for Pooling

Competitive binding assay. R + D1

• k1

k−1

• C1

. . . R + Di

• ki

k−i

• Ci

ITest−INC RTot

∝

Ka[L] 1+Ka[L]+Ka1[D1]+...

% Inhibition = IPC−ITest

IPC−INC × 100

=

Ka1[D1]+...+Kai[Di] 1+Ka[L]+Ka1[D1]+...+Kai[Di] × 100

Assume : All drugs present in equal & excess conc. Linear Model for Activity y = (1+Ka[L])

[D] %I 100−%I = P i Ki

y – modified measured quantity. Ka, [L] and [D] are known.

Linear Algebra Problem : y = MK

Motivation Pooling in HTS QUAPO Challenges Summary

QUAPO : Quantitative Analysis of Pooling in HTS

QUAPO

Sparsity : Most compound activities (Ka’s are close to zero (inactive). Linearity : Measured quantity maps linearly to compounds activity (with reasonable approximations). Solve min

x ||x||1 subject to ||Φx − y||2 ≤ ǫ

Motivation Pooling in HTS QUAPO Challenges Summary

Small Library Simulation

Synthetic Screen : small molecule ligands for formylpeptide receptor (FPR) with 6 showing activity.5 STD(n = 272, d = 3, e = 0%, r = 10) required m = 116 tests. y = (1+Ka[L])

[D] %I 100−%I = P i Ki

[L] = 1.5µM, 1/Ka = 3µM and [D] = 1.5µM

5Edwards et. al., Nature Protocols (2006)

Motivation Pooling in HTS QUAPO Challenges Summary

Small Library : QUAPO Result

Motivation Pooling in HTS QUAPO Challenges Summary

Challenge 1 : Pooling Design (Φ) Constraints

With existing HTS technology, easiest to use Sparse Binary Matrices (STD/DeVore matrix) or Expander Graphs. Mixing Constraint Compound concentration must be detectable in physiological range. Ionic strength of mixture must be low to prevent precipitation or changes to biological target. The assay must be reasonably simple to physically construct. Challenge 1 Row weight of Φ is tightly capped. Simple Heuristic : Not more than ∼ 10 compounds can be pooled in a test.

Motivation Pooling in HTS QUAPO Challenges Summary

Really Sparse Matrices Row weight cap implies that limited compression can be achieved.

Motivation Pooling in HTS QUAPO Challenges Summary

Challenge 2 : Liquid Handling Issue

Pooling at the level of individual compounds is hard and/or costly. Challenge 2 Original Library is subdivided into mutually exclusive blocks.

Motivation Pooling in HTS QUAPO Challenges Summary

Challenge 2 : A Simple Solution

Φ must be designed for smaller ˆ n and repeated in blocks on whole library n

Motivation Pooling in HTS QUAPO Challenges Summary

Challenge 3 : Measurement Error CS algorithms promise to handle additive noise. Small volumes and automation mean erasures are possible. Given Challenges 1 & 2, promising compression and error-correction might be difficult.

Challenge 3 Erasures of measurements are possible

Motivation Pooling in HTS QUAPO Challenges Summary

Challenge 4 : Non-additive behavior Synergy : pooled compounds react or aggregate to produce a hit Antagonism : pooled compounds block each other out Solution: Challenges can be treated as bugs or features. Bug : make designs more robust to these errors Feature : ability to detect mutli-compound drugs or drug cocktails

Challenge 4 Algorithms to handle non-additive behavior

Motivation Pooling in HTS QUAPO Challenges Summary

Advances in Pooling

Theme 6

Use chemical structure information about compounds while designing pools Simulations to predict probabilities of synergy or antagonism Simulations to evaluate average-case pooling design properties (theorems give worst-case bounds) Bayesian Decoders to evaluate various scenarios of compound interaction

6Will take more (compute) time

Motivation Pooling in HTS QUAPO Challenges Summary

Summary

Take Home Points

Current HTS strategies have hit a wall. Ever increasing compound collections and explosion of biological targets from genomics need a new approach. Age of multi-compound, multi-target therapeutics requires a paradigm shift in HTS. Pooling designs have the potential to be that change. Compressive Sensing can help make HTS quantitative (QUAPO). Lots of interesting (theory) problems need to be solved to make this approach practical. Currently implementing experimental validation at HTS facility in Univ. of Michigan.

Motivation Pooling in HTS QUAPO Challenges Summary

Acknowledgments At the University of Michigan : Peter Woolf Anna Gilbert Paul Shearer Systems Biology Group Center for Chemical Genomics

Questions . . . Comments . . . Suggestions

Thank You